User:Meng Xiao He/Notebook/fall08/2008/12/04

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  • set up new MR-1, wmDuo, and wdcco cultures for conjugation tomorrow

new cco candidate colonies

  • repatching of colonies from old cco plate: 1 colony was Gm sensitive, all were sucrose resistant
  • patching of colonies from new cco plate: all 20 were sucrose resistant and none were Gm sensitive, no obvious morphological diffs on plate, although liquid cultures had appeared to grow a little slow
  • Phusion PCR w/ Duo Xover

PCR redo

  • Previous PCR had high MW smears. Unclear if anything worked. Remainder of PCR mix run on 58C long cycle PCR to see if there will be high MW amplication corresponding to a complete, unrecombined out suicide vector.